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Novel coronavirus specific antigen peptide and application thereof

A technology of usage and homology, applied in the direction of viral antigen components, viral peptides, viruses, etc., can solve the problems of limited drugs and no effective vaccine reserves, etc., and achieve good accuracy and specificity

Active Publication Date: 2021-04-02
SUZHOU INST OF SYST MEDICINE +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for viral infectious diseases, the available drugs are very limited, and vaccines are one of the most effective means
However, due to the emergence and unpredictability of emerging infectious diseases, most emerging and severe infectious diseases do not have effective vaccine reserves

Method used

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  • Novel coronavirus specific antigen peptide and application thereof
  • Novel coronavirus specific antigen peptide and application thereof
  • Novel coronavirus specific antigen peptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1a

[0076] Synthesis of antigenic epitopes

[0077] We synthesized seven peptide epitopes that may have antibody responses, which are located at amino acids 25-39, 672-691, 764-778, 907-921, 1155-1169, 1198-1212, and 1257-1271 of the coronavirus S protein .

[0078] Synthetic method of antigenic peptide:

[0079] The peptide sequence was synthesized by GenScript Biotechnology, with a purity ≥ 85%, a gross weight of 4 mg, and 1 mg per 1 tube.

Embodiment 1b

[0081] Preliminary screening of specific antigenic peptides by ELISA

[0082] Enzyme-linked immunosorbent reaction Elisa experimental steps:

[0083] 1. MES powder (SIGMA, Lot #SLBZ3485) was prepared with ddH2O into 0.1 M MES buffer with pH=6.0; EDC (C8H17N3, Thermo Scientific, Lot #TB257918) was diluted with ddH2O to 10 mg / ml.

[0084] 2. Dilute the peptide to 4 μg / ml with 0.1M MES buffer.

[0085] 3. Set a negative control in the microplate, add 10μl EDC solution and 50μl peptide solution to each well; add 10μl EDC solution and 50μl MES buffer to the remaining wells. Shake gently to mix. Seal the plate with self-adhesive strips and store at 4°C overnight or at room temperature for more than two hours.

[0086] 4. Remove the adhesive strip, blot the liquid in the well, add 300μl ddH2O to each well, let it stand for 2 minutes, discard the liquid, and pat the plate dry. Repeat the above steps 2 more times.

[0087] 5. Prepare 1% BSA blocking solution with 1X PBST (10X PB...

Embodiment 2

[0097] The 3-D structure of SARS-CoV-2 S protein annotates B cell epitopes

[0098] Download the full genome sequence of SARS-CoV-2 (NC_045512) from NCBI GeneBank, and then extract the full-length sequence of the S protein for structural simulation. We submitted the S protein sequence on the SWISS-MODEL online server, obtained the SARS-CoV-2 S protein structure using the SARScoronavirus S protein (PDB ID: 6VSB) as a template, and performed epitope annotation on PYMOL.

[0099] figure 2 Represents the 3-D structure diagram of SARS-CoV-2S protein, protein S25-39, protein S672-691, protein S764-778, protein S907-921 are all in figure 2 is marked in.

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Abstract

The invention relates to a novel coronavirus specific antigen peptide and application thereof. Specifically, the invention relates to a polypeptide, which is an antigen peptide of SARS-CoV-2 virus, and application of the antigen peptide in disease diagnosis, preparation of a COVID-19 vaccine, and preparation of drugs for the prevention and treatment of COVID-19.

Description

technical field [0001] This disclosure belongs to the field of biotechnology. Specifically, the present disclosure relates to an antigenic peptide of SARS-CoV-2 virus, and the use of the aforementioned antigenic peptide in the preparation of COVID-19 vaccines, drugs for the prevention and treatment of COVID-19. Background technique [0002] The novel coronavirus (SARS-CoV-2) belongs to the Betacoronavirus genus and is a linear single-stranded RNA (ssRNA) virus. The full length of its genome is about 29903 nucleotides, including 10 genes. Since the first SARS-CoV-2 genome sequence data was released on January 10, 2020, several genome sequences of novel coronaviruses isolated from patients have been released successively since then. On January 22, 2020, the Genome Science Data Center officially released the 2019 Novel Coronavirus Resource Library. According to data analysis, the genome sequence similarity between the 2019 novel coronavirus (SARS-CoV-2) and the SARS virus th...

Claims

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Application Information

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IPC IPC(8): C07K14/165G01N33/68G01N33/569A61K39/215A61P31/14
CPCC07K14/005G01N33/6854G01N33/56983A61K39/12A61P31/14C12N2770/20022C12N2770/20034G01N2333/165G01N2469/20
Inventor 杨衡秦成峰高美玲王雅楠邓永强
Owner SUZHOU INST OF SYST MEDICINE
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